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DIVERSIDADE GENÉTICA DE ACESSOS DO BANCO ATIVO DE GERMOPLASMA DE Vitis Spp

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DIVERSIDADE GENÉTICA DE ACESSOS DO BANCO ATIVO DE GERMOPLASMA DE Vitis Spp INSTITUTO AGRONÔMICO PROGRAMA DE PÓS-GRADUAÇÃO AGRICULTURA TROPICAL E SUBTROPICAL DIVERSIDADE GENÉTICA DE ACESSOS DO BANCO ATIVO DE GERMOPLASMA DE Vitis spp. DO INSTITUTO AGRONÔMICO GEOVANI LUCIANO DE OLIVEIRA Orientadora: Dra. Mara Fernandes Moura Coorientadora: Dra. Lívia Moura de Souza Dissertação apresentada como requisito para a obtenção do título de Mestre em Agricultura Tropical e Subtropical, Área de Concentração em Genética, Melhoramento Vegetal e Biotecnologia. Campinas, SP 2020 Ficha elaborada pela bibliotecária do Núcleo de Informação e Documentação do Instituto Agronômico O48d Oliveira, Geovani Luciano de Diversidade genética de acessos do banco ativo de germoplasma de Vitis spp. do Instituto Agronômico / Geovani Luciano de Oliveira. Campinas, 2020. 101 fls. Orientadora: Mara Fernandes Moura Co-orientadora: Lívia Moura de Souza Dissertação (Mestrado) Agricultura Tropical e Subtropical – Instituto Agronômico 1. Melhoramento genético. 2. Coleção nuclear . 3. Recursos genéticos 3. Marcador molecular 4. SSR. I. Moura, Mara Fernandes II. Souza, Lívia Moura de III. Título CDD. 574.182 ii DEDICO A Deus e a minha família por serem minha base. Em especial aos meus pais, Adelar Bueno de Oliveira e Eva Maria de Oliveira. iii AGRADECIMENTOS Primeiramente a Deus, por ter me dado saúde, força e ânimo durante essa etapa e por ter permitido mais esta vitória. Aos meus pais Adelar e Eva, pelo amor, incentivo е apoio incondicional. Agradeço por toda ajuda que me deram durante a realização deste trabalho. Ao meu irmão Gustavo, pelo companheirismo e apoio durante toda a vida. A minha namorada Bianca, por acreditar em mim e em meu potencial, pela compreensão, carinho e atenção. Obrigado por ser essa pessoa incrível, agradeço a Deus todos os dias por ter você em minha vida. À Dra. Mara F. Moura, pela orientação, dedicação, conselhos, incentivo, amizade e sobretudo pela oportunidade de realizar um trabalho gratificante e enriquecedor à minha formação profissional. À Dra. Anete Pereira de Souza por acreditar neste trabalho, por todo apoio estrutural e por me receber como aluno de Doutorado. À Dra. Lívia M. de Souza, pela oportunidade de ser seu coorientado, por acreditar no meu trabalho e por todo auxílio prestado. À Dra. Fernanda A. de Oliveira, pelo apoio e dedicação durante o desenvolvimento deste trabalho. Obrigado pela paciência e pela imensa ajuda. A todos os colegas do Laboratório de Análise Genética e Molecular (LAGM), pelos esclarescimentos de dúvidas e auxílio nas análises. À Dra. Maria I. Zucchi, pelos conselhos e pela grande ajuda nas análises. Ao programa de Pós-graduação do IAC, pela oportunidade e por todo suporte. A CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) pela concessão da bolsa durante o período do mestrado. À FAPESP (Fundação de Apoio à Pesquisa do Estado de São Paulo) pelo apoio financeiro ao projeto (processo: 2018/13539-9). Aos meus amigos Gustavo, Isabella e Cínthia, por toda ajuda, conversas, risadas, companheirismo e pelos agradáveis momentos vividos. A todos que, mesmo não tendo citados aqui, contribuíram de alguma forma para a conclusão dessa etapa. Muito obrigado! iv SUMÁRIO LISTA DE TABELAS .............................................................................................................. vi LISTA DE FIGURAS ............................................................................................................. viii RESUMO ................................................................................................................................... x ABSTRACT .............................................................................................................................. xi 1. INTRODUÇÃO ...................................................................................................................... 1 2. REVISÃO BIBLIOGRÁFICA .............................................................................................. 4 2.1 O Gênero Vitis ...................................................................................................................... 4 2.2 Importância econômica ........................................................................................................ 5 2.3 Programas de melhoramento genético da videira ................................................................. 7 2.4 Diversidade genética............................................................................................................. 9 2.5 Marcadores moleculares no melhoramento genético ......................................................... 10 3. MATERIAL E MÉTODOS .................................................................................................. 14 3.1 Material vegetal .................................................................................................................. 14 3.2 Coleta de folhas e extração do DNA .................................................................................. 24 3.3 Seleção dos locos microssatélites e condições de amplificação ......................................... 25 3.4 Caracterização molecular do Banco Ativo de Germoplasma ............................................. 27 3.5 Análise estatística dos dados .............................................................................................. 27 3.5.1 Análise da diversidade genética ...................................................................................... 27 3.5.2 Estrutura genética ............................................................................................................ 28 3.5.3 Amostragem da coleção nuclear (Core collection) ......................................................... 29 3.6 Análise de genealogia do híbrido SR 501-17 ..................................................................... 29 4. RESULTADOS E DISCUSSÃO ......................................................................................... 30 4.1 Caracterização do germoplasma ......................................................................................... 30 4.2 Diversidade genética........................................................................................................... 43 4.2.1 Relação genética e estruturação dos acessos ................................................................... 48 4.3 Coleção nuclear (Core collection) ...................................................................................... 58 4.4 Identificação da genealogia do híbrido SR 501-17 ............................................................ 60 5. CONCLUSÕES .................................................................................................................... 62 6. CONSIDERAÇÕES FINAIS ............................................................................................... 63 7. REFERÊNCIAS ................................................................................................................... 64 ANEXO 1. ................................................................................................................................ 76 ANEXO 2. ................................................................................................................................ 89 v LISTA DE TABELAS Tabela 1 - Principais países produtores de uvas de mesa, uvas passas e uvas para vinho nos anos de 2014 a 2018. Fonte: OIV (2019) ..................................................... 6 Tabela 2 - Acessos do BAG de Vitis spp. do Instituto Agronômico (IAC) classificados quanto ao táxon, origem, parentais e uso. ......................................................... 15 Tabela 3 - Relação dos 17 marcadores microssatélites utilizados para caracterizar os acessos do BAG de Vitis spp. do Instituto Agronômico (IAC), com respectivos grupos de ligação, motivos de repetição e referências. ..................................... 26 Tabela 4 - Acessos do BAG de Vitis spp. do Instituto Agronômico (IAC) precisamente identificados por meio de correspondência a um perfil SSR de referência validado. ............................................................................................................ 31 Tabela 5 - Acessos do BAG de Vitis spp. do Instituto Agronômico (IAC) que tiveram seus perfis genéticos correspondidos com perfis genéticos referência de nomes diferentes. .......................................................................................................... 35 Tabela 6 - Acessos do BAG de Vitis spp. do Instituto Agronômico (IAC) que não tiveram seus perfis genéticos correspondidos a qualquer perfil de referência validado 37 Tabela 7 - Duplicatas encontradas no BAG de Vitis spp. do Instituto Agronômico (IAC) através da análise de 17 locos microssatélites. ................................................. 40 Tabela 8 - Acessos sinônimos encontrados no BAG de Vitis spp. do Instituto Agronômico (IAC) por meio da análise de 17 locos microssatélites. .................................... 42 Tabela 9 - Estimativas de parâmetros genéticos dos 17 locos microssatélites analisados em 410 acessos pertencentes ao BAG de Vitis spp. do Instituto Agronômico (IAC). ........................................................................................................................... 45 Tabela 10 - Estimativas de parâmetros de diversidade genética dos acessos do BAG de Vitis spp. do Instituto Agronômico (IAC) considerando os grupos identificados pelo STRUCTURE em K=3 e os acessos admixture (q < 0,7 Grupo 4). ................. 51 Tabela 11 - Análise de variância molecular
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